CN104523591A - Formula and preparation method of non-allergenic and painless novel propofol fatty microemulsion freeze-drying preparation - Google Patents
Formula and preparation method of non-allergenic and painless novel propofol fatty microemulsion freeze-drying preparation Download PDFInfo
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Abstract
The invention discloses a formula and a preparation method of a non-allergenic and painless novel propofol fatty microemulsion freeze-drying preparation. The novel preparation comprises artificially synthesized phospholipid and artificially synthesized fatty acid glyceride, the presence of such allergic sources as a trace of protein in lecithin and soybean oil as natural sources is avoided, the uses of a metal complex EDTA, an antioxidant sulfite and thiosulfate is avoided, the allergic reaction caused by a metal complexing agent and sulfide is eliminated; a water phase free propofol is removed through various means in the formulation and preparation process, so that the injection pain is alleviated or eliminated and thus the safety of the preparation and the compliance of patients are improved.
Description
Technical field
The invention belongs to medical drugs series, relate to a kind of formula and preparation of lyophilized formulations of medicine, particularly relate to the formula without sensitization, painless novel propofol fat micro emulsion frozen preparation and preparation.
Background technology
Propofol (also known as dipropofol, propofol, 2,6-Bis(1-methylethyl)phenol, structure is shown in Fig. 1) and be a kind of hindered phenol, lipotropy is strong, is used for clinical anesthesia induction and maintains mainly with fat emulsion formulation dosage form
[1].Propofol is rapid-action with it as a kind of clinical conventional intravenous anesthetic, it is short to hold time, revive rapidly, metabolism is fast, be widely used without the feature that accumulation, safety are high
[2].
Propofol injection is along with it uses increasingly extensive clinically, and the report of its untoward reaction increases increasingly
[3].Propofol injection untoward reaction complicated clinical manifestation is various, and the ratio that anaphylactic shock occurs is higher
[4], seriously can cause death.The description points for attention of propofol injection (Jing'an) are mentioned: this product is containing soybean oil, and serious anaphylaxis may appear in only a few patient.And untoward reaction is also pointed out: hypotension and of short duration respiration inhibition may be occurred.There is anaphylaxis after using propofol in only a few case, comprises QuinckeShi edema, bronchospasm, erythema and hypotension.The pharmaceutical control and administration department of various countries comprising U.S. FDA is all clearly indicated in the description of the propofol injection of approval: to the allergies such as Semen sojae atricolor, egg yolk, person prohibits the use.In addition, propofol Injection administration process can cause injection site incidence rate up to the injection pain of (30-70%), causes have a strong impact on the life of patient, body and spirit
[5-7], thus limit it and clinically further to apply.Therefore exploitation is without sensitization, and painless novel propofol formulations has certain Clinical practice to be worth.
The clinical practice dosage form of current propofol is lipomul, and usually adopt natural yolk lecithin and soybean oil as adjuvant, they contain micro-yolk protein and soybean protein, easily cause anaphylaxis.These adjuvants or good natural medium, support microbial growth.If improper use, easily cause postoperative infection, lead to grave consequences
[8].Propofol the U.S. listing after 1 year between, said preparation constantly has the report causing postoperative infection and heating
[9-11].In 2010, FDA (Food and Drug Adminstration) (FDA) represented, the problem due to production aspect forces ladder watt to suspend with Hospira Inc (HSP) and produces its propofol sedation agent, and recalls portioned product.In order to prevent the generation of postoperative infection, add EDTA and sulphite in regular dosage form as antimicrobial component.
In addition, in propofol structure, there is phenolic hydroxyl group, easy oxidized generation 2,6-diisopropyls-Isosorbide-5-Nitrae-benzoquinone.Also can polymerization reaction take place and generate 3,3,5,5 '-tetra isopropyl xenol (chemical constitution is shown in Fig. 2), these two impurity have larger toxicity
[12]; Moreover many unsaturated double-bonds compound in adjuvant, easily there is lipid peroxidation in unstable chemcial property, produces peroxide in atmosphere
[2].In order to anti-oxidation, conventional formulation adds sulphite usually, or thiosulfate, or adds EDTA and carry out complexation of metal ions, prevent the oxidation of propofol and adjuvant, but EDTA and sulphite can cause allergic reaction
[13], especially to essential asthma patient, the propofol of application containing EDTA or sulphite age>=40, long-term smoking patient can cause total pulmonary resistance to increase.
In order to reach the effect without sensitization, in novel formulation of the present invention, not using EDTA, sulphite or thiosulfate, effectively preventing anaphylactoid generation.In addition in order to still ensure the stability of preparation under the situation not adding above adjuvant, we have been made into fatty micro emulsion frozen preparation, and use brown cillin bottle as inner packing.Phenolic hydroxyl group is there is in propofol structure, easily oxidized.Illumination can accelerated oxidation reaction generation.Mostly be unsaturated double-bond compound in adjuvant in addition, chemical property is also unstable.Brown cillin bottle can accomplish hypersorption at 200nm ~ 400nm, and absorption curve enters visible-range
[14-15].Illumination effectively can be avoided propofol and adjuvant prooxidant effect, increase preparation stability.Test proves, use brown cillin bottle after being prepared into micro emulsion frozen preparation, the stability of sample increases and effectively prevent the infection of the microorganisms such as antibacterial, also to prevent the light degradation reaction generation of preparation, specifically sees embodiment.
Propofol Injection pain has been classified as the topmost side effect of propofol by anesthesia circle and medicine administrative organ.In order to alleviate, suppressing propofol Injection pain, people have carried out large quantity research to propofol Injection Theory of Pain Mechanism, and the result of study of comprehensive injection pain mechanism at present, injection pain inducement can be summarized as three hypothesis: 1, free propofol directly stimulates hypothesis
[16-18]; 2, free propofol indirectly excites and produces Kallidin I hypothesis
[19-21]; 3, free propofol causes prostaglandin release hypothesis
[22].Propofol vein injection causes injection pain to be a complicated process, from current most result of study analysis, that is a kind of cause injection pain mechanism all with aqueous phase in and the propofol that sticks of emulsion droplet outer surface have direct relation.Not accommodating doubtful, the propofol that in pharmaceutical preparation aqueous phase, free propofol and emulsion droplet outer surface stick is the arch-criminal causing pain.
For the untoward reaction such as anaphylaxis and injection pain of conventional formulation, we have done following improvement.1, adopt synthetic phospholipid and synthetic fatty glyceride, fundamentally eliminate the existence in the sensitization sources such as trace in natural phosphatidyl choline in conventional formulation and soybean oil.2, do not use metal complex EDTA, be not used as sulphite or the thiosulfate of antioxidant yet, eliminate the anaphylaxis that metal chelating agent and sulfide may cause.3, adopt the form of fatty micro emulsion frozen preparation, make preparation more stable.Solve a difficult problem easily oxidized under not having anti-oxidant protective agent, effectively prevent the oxidative degradation of propofol and adjuvant; Also effectively prevent the growth of preparation antibacterial in preservation process simultaneously.4, adopt brown cillin bottle as interior packaging material, avoid the degraded of the propofol caused because of illumination, and light-catalysed phospholipid oxidation
[2].5, advanced technologies is adopted, effectively reduce and be not wrapped in propofol free in fatty microemulsion emulsion droplet or the propofol in the absorption of emulsion droplet outer surface, and envelop rate does not reduce after redissolving, free propofol aqueous phase content≤10 μ g/ml, particle diameter is less than 180nm and meets the requirements, reduce free propofol to the stimulation of blood vessel, thus reach without injection pain.
The present invention is by avoiding using possible sensitization source in prescription, adopt lyophilized formulations, brown bottle has ensured product stability during preservation, and adopts the independent of multiple method and conbined usage to reach minimizing or the basic object removing the propofol that free propofol and emulsion droplet outer surface stick in the rear aqueous phase of micro emulsion frozen preparation redissolution.Owing to fundamentally eliminating anaphylactogen and avoiding the induced pain root causing pain, prepared propofol fat micro emulsion frozen preparation reaches without sensitization, painless or substantially painless ideal effect, becomes the better preparation of safety.
Three, summary of the invention
The object of the present invention is to provide a kind of without sensitization propofol fat micro emulsion frozen preparation, composed of the following components:
Described synthetic phospholipid is characterized in that described synthetic phospholipid is the mixing of one or more of formula I structure;
R
2:-CH
2-CH
2-N
+H
3(PE)
-CH
2-CH
2-N
+(CH
3)
3(PC)
-C
6H(OH)
5(PI)
-CH
2COO
-CH(N
+H
3) (PS)
-CH
2CHOH-CH
2OH (PG)
Wherein, R
2represent and be selected from choline (PC), ethanolamine (PE), inositol (PI), glycerol (PG), serine (PS), R and R
1can be identical or different, be selected from saturated, cholesterol or polyunsaturated in, long chain hydrocarbon groups, as alkyl, thiazolinyl, alkynyl, R and R
1length optional 4 ~ 26 carbon of carbochain, the length of preferred carbochain is 6 ~ 22 carbon, and more preferably the length of carbochain is 10 ~ 20 carbon.Can be synthetic dipalmitoyl phosphatidyl choline (DPPC), synthetic distearoyl phosphatidylcholine (DSPC), synthetic dilinoleoylphosphatidylcholine, synthetic linolenic acid phosphatidylcholine, synthetic DOPC; The sub-oleolyl phosphatidyl choline of synthetic 1-stearoyl-2 arachadoyl phosphatidyl choline, synthetic 1-palmityl-2-, synthetic and stearyl glycerol (DSPG), synthetic distearyl ethylethanolamine etc. one or more.
Described synthetic fatty glyceride is one or more mixing of formula II;
Wherein, R, R
1and R
2for can be identical or different, be selected from saturated, cholesterol or polyunsaturated middle long chain hydrocarbon groups, as alkyl, thiazolinyl, alkynyl, R, R
1and R
2length optional 4 ~ 26 carbon of carbochain, the length of preferred carbochain is 6 ~ 22 carbon, and more preferably the length of carbochain is 10 ~ 20 carbon; Can be one or more of glyceryl linoleate etc. of synthetic tristerin, synthetic tripalmitin, synthetic olein, synthetic glyceryl linolenate, synthetic.
Described synthetic mono fatty acid glyceride is one or more mixing of formula III;
Wherein, R is selected from saturated, cholesterol or polyunsaturated middle long chain hydrocarbon groups, and as alkyl, thiazolinyl, alkynyl, the length of the carbochain of R is 16 ~ 18 carbon.Can be one or more of glyceryl monostearate, monopalmitin, glyceryl monolaurate, glyceryl monooleate etc.
Described polyethyleneglycol lipid derivates can be Polyethylene Glycol-DSPE, PEG-CHOL, one or more of Polyethylene Glycol-fatty acid etc.
Described skeletal support agent can be one or more in lactose, sucrose, maltose, mannitol, sorbitol, xylitol.
Preferably, the weight ratio of propofol and synthetic fatty glyceride is less than 1:15, raising oil phase contributes to controlling propofol relative to the ratio of propofol in oil phase diluent, is in relatively low concentration (adopting relatively large oil to carry out dissolved substance), makes propofol more effectively and fully can be dissolved in oil phase and be wrapped in fatty microemulsion emulsion droplet.
More preferably, the weight ratio of propofol, synthetic fatty glyceride and synthetic phospholipid is 1:25:(0.8 ~ 3.2), improve phospholipid ratio, under the prerequisite of proportions constant keeping oily diluent and propofol, improve the consumption of phospholipid, thus ensure to improve the parcel efficiency of propofol in fatty microemulsion.
Propofol fat micro emulsion frozen preparation of the present invention, preparation method is as follows:
Whole process is carried out under nitrogen protection.Get heated and stirred, the mixings such as the propofol of recipe quantity, synthetic phospholipid, synthetic fatty glyceride and Polyethylene Glycol phospholipid derivative; separately get appropriate water for injection; add oleic acid and skeletal support agent dissolving; under nitrogen protection condition; oil solution is added slowly in the aqueous phase of cutter high-speed stirred (8000rpm); continue 15min, obtain colostrum solution.Through high pressure homogenizer homogenizing 7 ~ 8 times, pressure 100MPa, regulate pH6.0-8.0, control particle diameter and be less than 180nm, adopt the independent of multiple method and conbined usage to reach and reduce or substantially remove the propofol that in the rear aqueous phase of micro emulsion frozen preparation redissolution, free propofol and emulsion droplet outer surface stick, filter, fill, prepare lyophilized formulations: at subzero less than 45 DEG C pre-freeze 2 ~ 5h, be warmed up between subzero 30 DEG C to subzero 15 DEG C and maintain more than 22h, vacuum decompression removing moisture, be warming up to 0 ~ 5 DEG C, maintain 5h and continue vacuum drying decompression removing moisture, be warming up to 25 DEG C, continue vacuum decompression and go out moisture, the freeze-dried emulsion of drying,
Preferably, after fatty microemulsion homogenizing completes, use ultrafiltration or dialysis process to remove propofol free in aqueous phase, in step, select brown cillin bottle fill.
The technique that the free propofol of preparation aqueous phase is removed in ultrafiltration is shown in Fig. 2.Get propofol fat microemulsion 1000mL (Specific amounts according to the design of process units, and effective process capacity of ultrafiltration post is different and change), take a morsel its aqueous phase free propofol concentration of sample analysis.According to Fig. 2 assembling " ultrafiltration production equipment ", select the ultrafiltration post core in the aperture be applicable to, the principal element of selection can freely be passed through with aqueous solution, and fat milk is by being not Selecting All Parameters.First isotonic buffer solution equilibria ultrafiltration post is used, then slowly by propofol fat microemulsion input ultrafiltration post, adjustable column pressure.Aqueous solution containing free propofol, through ultrafiltration post, is effectively separated with propofol fat microemulsion.The solution of disappearance is supplemented by buffer solution.Generally exchanged by the buffer solution of 10 times of volumes and the free propofol in aqueous phase just can be ensured all to remove totally.Concrete technological parameter, preparation disposal ability, buffer solvent exchange capacity must be determined according to the control overflow of the free propofol content in concrete equipment and final preparation.
It should be noted that, the present invention adopts 1), use artificial synthetic phospholipid and synthetic fatty glyceride in prescription, fundamentally go the existence in the sensitization source such as lecithin and soybean oil trace albumin avoiding natural origin.2), do not use metal complex EDTA, sulphite or thiosulfate sulfides, eliminate the existence of the anaphylactogen such as metal chelating agent and sulfide.3), adopt the form of fatty micro emulsion frozen preparation, make preparation more stable, effectively prevent the oxidative degradation of propofol and adjuvant and the bacterial growth in preservation process.4), adopt brown cillin bottle as interior packaging material, avoid the degraded of the propofol caused because of illumination, and light-catalysed phospholipid oxidation.5), being used alone or in combination of following Lung biopsy is adopted, decrease the free propofol that micro emulsion frozen preparation redissolves in rear aqueous phase, or the content of the propofol that fatty microemulsion outer surface sticks, after redissolving, the envelop rate of propofol does not reduce, free propofol content≤10 μ g/ml in aqueous phase, and particle diameter is less than 180nm meets the requirements, reduce free propofol to vascular stimulation, without injection pain.
The Lung biopsy that the present invention describes is: 1), fatty microemulsion is homogeneous complete after, use ultrafiltration (see Fig. 3), dialysis or additive method to remove free propofol in aqueous phase.2) ratio of oily diluent and propofol, is improved, namely control propofol in oil phase diluent, be in relatively low concentration (adopting relatively large oil to carry out dissolved substance), make propofol more effectively and fully can be dissolved in oil phase and be wrapped in fatty microemulsion emulsion droplet.3), improve phospholipid ratio, under the prerequisite of proportions constant keeping oily diluent and propofol, improve the consumption of phospholipid, thus ensure to improve the parcel efficiency of propofol in fatty microemulsion.4) in formula, add Polyethylene Glycol phospholipid derivative (form structure and see schematic diagram 4).Polyethylene Glycol phospholipid derivative both can strengthen emulsifying effectiveness, ensured that propofol can effectively be wrapped in fatty microemulsion oil droplet; Can drip at fatty microemulsion again and form polymeric buffer band (Fig. 5 is shown in physical barrier effect) between blood vessel wall, common fats breast propofol is very near blood vessel endothelium surface, and the propofol of fatty emulsion droplet outer surface and vessel wall contact can produce to stimulate or cause inflammation.And Polyethylene Glycol fatty micro emulsion frozen preparation is redissolved after away from blood vessel wall, the propofol on emulsion droplet surface not with vessel wall contact, so stimulation can not be produced, thus without injection pain.5), employing disaccharide etc. are as the skeletal support agent of lyophilized formulations.Disaccharide etc. are except the isotonic agent function of itself, they can reduce and are attached on the propofol that fatty microemulsion drips outer surface in fatty microemulsion propofol forming process, the parcel efficiency of propofol in fatty microemulsion can be improved, in addition disaccharide etc. can also form physics isolation strip between fatty microemulsion emulsion droplet and blood vessel wall, and what avoid free propofol and blood vessel wall touches building.
The novel propofol fat micro emulsion frozen preparation being combined preparation by above-mentioned all method or Part Methods can reduce or eradicate propofol fat emulsion formulation injection stimulation and injection pain, improves the toleration of patient, has very important clinical meaning.
The present invention is above without sensitinogen adjuvant and painless solution by adopting simultaneously, reaches the novel novel propofol fat micro emulsion frozen preparation without sensitization, nothing injection pain of preparation.
Concrete without sensitization, prepare embodiment without the novel propofol fat micro emulsion frozen preparation of pain
In order to clearer elaboration innovative point of the present invention, below classification provides without sensitization, the present invention is described in further detail to prepare embodiment without the novel propofol fat micro emulsion frozen preparation of injection pain.
Important analysis method
(1) aqueous phase propofol content detection method after, redissolving:
After propofol fat micro emulsion frozen preparation redissolves, in aqueous phase, the mensuration of propofol content measures after centrifuging can be adopted aqueous phase, oil phase and emulsifying oxidant layer to be separated respectively.Utilize in this experiment and centrifugal emulsifying oxidant layer to be effectively separated with aqueous phase, then adopt HPLC to determine fatty microemulsion aqueous phase drug content:
The injection that precision measures after the redissolution of propofol fat micro emulsion frozen preparation is appropriate, make the sample solution obtaining 50ug/ml in measuring bottle, add Chromatographic Pure Methanol to scale, mixing, 0.22 μm of filtering with microporous membrane, get subsequent filtrate, measure by chromatographic condition HPLC method under assay item, external standard method calculates preparation of Chinese medicine total concentration Ctotal; The injection separately got after the redissolution of propofol fat micro emulsion frozen preparation is about 10mL, put in supercentrifuge with 30, the centrifugal 1.0h of 000rpm, temperature 4 DEG C, take off layer settled solution through 0.22 μm of filtering with microporous membrane, get subsequent filtrate, measure (Jasco PU-2089 type quaternary pump high performance liquid chromatograph by chromatographic condition HPLC method under assay item, Jasco-UV-2075plus detector), external standard method calculates aqueous phase drug concentration Cwater.
Chromatographic condition: chromatographic column: Kromasil-C184.6 × 250mm, 5 μm; Mobile phase: acetonitrile-water (85:15); Determined wavelength: 280nm; Flow velocity: 1.0ml/min; Sample size: 20 μ l; Column temperature: 25 DEG C.
(2), granularity and particle size distribution method
A. instrument
Zeta potential/particle Sizer (Nicomp
tM380ZLS, U.S. PSS.NICOMP).
B. algoscopy
Get this product appropriate, redissolve with 0.9% sodium chloride injection, and dilute 5000 times, mixing, as need testing solution, measure by Dynamic laser scattering particle size determination method (Chinese Pharmacopoeia version in 2010 two annex Ⅸ E the 3rd methods), the particle diameter that microemulsion drips should between 10 ~ 180nm, and the granularity 99% that microemulsion drips should below 0.2 μm.
(3), propofol fat micro emulsion frozen preparation anaphylaxis inspection technique
A. principle
A certain amount of need testing solution injects in Cavia porcellus body by this genealogy of law, and separated in time posterior vein injection need testing solution excites, and observes animal and occurs anaphylactoid situation, to judge whether test sample causes animal systemic anaphylaxis.
On approbation Cavia porcellus should be healthy qualified, and body weight 250-350g, female Mus should without pregnant.Before the test with in process of the test, all should raise by normal rearing conditions.The Cavia porcellus doing this test must not be reused.
B. reagent
Diprivan
tMinjection (commercially available), propofol fat micro emulsion frozen agent (self-control), saline control group (commercially available), 5% Ovum Gallus domesticus album normal saline.
C. inspection technique
Get above-mentioned Cavia porcellus, be divided into 6 groups at random, often organize 6.The next day every only each lumbar injection need testing solution 1ml, totally 3 times, after final injection 10 days, by hind leg saphena respectively injection 2.0ml/ only, carry out sensitization.Observe behavior and the sign of every animal every day, first sensitization and weigh before exciting and record the body weight of every animal.Observing attack to excite in latter 30 minutes animal with or without symptoms of allergic.
D. result judges
Animal anaphylaxis progression is calculated by table 1 requirement.When the order of reaction is below 2 grades, then think qualified by the hypersensitive test of test product.
Table 1 Cavia porcellus anaphylaxis grading table
(4), propofol fat micro emulsion frozen preparation vascular pain evaluation test
Instrument, reagent and animal
A. instrument
Electromyogram monitor (AB-621G, Nihon Kohden Corp.), constant temperature blender with magnetic force (Shanghai Zhi Guang instrument and meter company limited), 100000/electronic analytical balance (BP211D Germany Sai Duolisi), the coaxial needle electrode of electromyogram and reference electrode (indifferent electrode), digital display thermostat water bath (Fuhua Instrument Ltd. of Jintan City).
B. reagent
Diprivan
tMinjection (commercially available), propofol fat micro emulsion frozen agent (self-control), blank fat milk (self-control), normal saline (commercially available).
C. animal
SD rat, weight 200 ~ 250g is male, is provided by The Fourth Military Medical University's Experimental Animal Center.
D. experimental technique
First male SD rat is divided into 6 groups, often organize 6, etherization, be fixed on Mus platform to face upward appearance, except right rear leg field of operation and electrode insert position hair, expose femoral artery, by polyethylene catheter intubate in Rat Right femoral artery, A/C simultaneously, electromyogram coaxial needle electrode and reference electrode are placed in right rear leg, and are connected to electromyogram monitor.
Give propofol fat milk sample product that commercially available and embodiment prepared the voluntarily femoral artery to rat respectively, evaluated the degree of vascular pain by the electromyography at the contiguous position of the tremulous pulse of administration sample.Electromyography method evaluation vascular pain is according to list of references method
[23].Electromyography record is started before administration propofol fat sample.To Post operation 1 hour, electromyography waveform stabilization, by intubate respectively the commercially available propofol injection of administration, the fatty microemulsion removing free propofol, highly dissoluble oil propofol fat microemulsion, improve the 1-27 embodiment sample such as propofol fat micro emulsion frozen preparation of the ratio of oil for injection, record electromyogram respectively after administration, and calculate area under the peak in electromyogram.
After the commercially available propofol injection of administration 1 hour, the fats emulsion sample (each 0.5ml) of embodiment regulated pH to 8, to calculate in electromyography waveform area under peak.The value measuring each group of every rat compares with reference to medicine with commercially available, to determine its " electromyogram area ratio ", represents with (%), represents the index of vascular pain.When this index is less than 100%, it shows that vascular pain alleviates compared with commercially available propofol fat emulsion injection.And index (electromyogram area ratio) is less, it is better that vascular pain alleviates effect.
(5) EDTA, sulphite, the propofol fat micro-emulsion injecta of thiosulfate and comparing of micro emulsion frozen preparation germ contamination is not added
Experiment grouping: experiment component is three groups: A group is the propofol fat microemulsion injection not adding EDTA, sulphite, thiosulfate; B group is the propofol fat micro emulsion frozen preparation not adding EDTA, sulfate; C group is for using physiological saline solution.
The preparation of need testing solution: under non-sterile conditions, according to formula preparation propofol fat microemulsion solution, gets and a certain amount ofly prepares propofol fat microemulsion injection, separately get equivalent solution and prepare fatty micro emulsion frozen preparation.The contrast of total number of bacteria is carried out after lyophilizing completes.Get appropriate propofol fat microemulsion injection respectively and fatty micro emulsion frozen preparation after redissolving, with physiological saline solution 10 times of gradient dilutions.
The comparison of clump count: get 9 clean glass dishes, inoculate 1ml A respectively, redissolve after B solution, dilution 10
2, 10
4, 10
6the B solution of A doubly and equal extension rate and C, shake up in heats liquefied nutrient agar, cultivates 24 hours, compare the clump count in culture medium after cooled and solidified at 37 DEG C.
Accompanying drawing explanation
The chemical constitution of Fig. 1, propofol
The oxidation of Fig. 2, propofol and polymerization impurity
A:2,6-diisopropyl-Isosorbide-5-Nitrae-benzoquinone
B:3,3,5,5 '-tetra isopropyl xenol
The process schematic representation of the free propofol after Fig. 3, ultrafiltration removal propofol fat micro emulsion frozen preparation redissolve in aqueous phase.
Schematic diagram after Fig. 4, polyethyleneglycol modified propofol fat micro emulsion frozen preparation redissolve.
Fig. 5, polyethyleneglycol modified propofol fat micro emulsion frozen preparation effectively alleviate the action principle schematic diagram of injection pain.
Fig. 5 A: common fats breast propofol
Fig. 5 B: polyethyleneglycol modified propofol fat micro emulsion frozen preparation
Under the Electromyographic peak of Fig. 6, invention laboratory sample method 1-4 area and commercial samples electromyogram peak under the ratio of area.
1), the sample (embodiment 1) prepared of hyperfiltration process;
2) sample (embodiment 14) prepared by phospholipid consumption, is increased;
3) sample (embodiment 24), using polyethyleneglycol lipid derivates to prepare;
4) sample (embodiment 10) of oil preparation at high proportion, is used.
Detailed description of the invention
Method one, use ultrafiltration, dialysis or additive method remove the investigation of the free propofol redissolved in rear aqueous phase.
Embodiment 1: ultrafiltration is investigated the suitability of common propofol fat microemulsion
Get propofol fat microemulsion 1000mL prepared by any one embodiment, take a morsel its aqueous phase free propofol concentration of sample analysis.According to Fig. 2 assembling " ultrafiltration production equipment ", select the ultrafiltration post core being applicable to aperture, the principal element of selection is with aqueous solution, and the free propofol particularly in aqueous phase can freely pass through, and fat milk is by being not Selecting All Parameters.First use isotonic buffer solution equilibria ultrafiltration post, then slowly propofol fat emulsion is inputted ultrafiltration post, adjustable column pressure.Aqueous solution containing free propofol, through ultrafiltration post, is effectively separated with propofol fat microemulsion.The solution of disappearance is supplemented by buffer solution.Generally exchanged by the buffer solution of 10 times of volumes and the free propofol in aqueous phase just can be ensured all to remove totally.
Embodiment 2: ultrafiltration is investigated the suitability of Polyethylene Glycol propofol fat microemulsion
Get the Polyethylene Glycol propofol fat microemulsion sample 1000mL prepared according to embodiment 21-24, take a morsel and analyze its aqueous phase free propofol concentration.According to Fig. 2 assembling " ultrafiltration production equipment ", selecting the ultrafiltration post core in the aperture be applicable to, can freely pass through with aqueous solution, and fat milk is by being not Selecting All Parameters.First use isotonic buffer solution equilibria ultrafiltration post, then slowly propofol fat emulsion is inputted ultrafiltration post, adjustable column pressure.Aqueous solution containing free propofol, through ultrafiltration post, is effectively separated with propofol fat microemulsion.The solution of disappearance is supplemented by buffer solution.Generally exchanged by the buffer solution of 10 times of volumes and the free propofol in aqueous phase just can be ensured all to remove totally.
Ultrafiltration can not affect the stability of Polyethylene Glycol propofol fat microemulsion, also can not cause Polyethylene Glycol coming off from fatty microemulsion surface.
The synthetic supplementary material of method two, different proportion is to the investigation of propofol content free in aqueous phase after redissolution.
(1), the proportion of propofol and flux oil is investigated
The usage ratio scope of the present invention to listed propofol and flux oil has been carried out verifying and has been investigated.Under technology and condition of the present invention, keep the constant rate of other adjuvant, the agent of the claims in the present invention propofol micro emulsion frozen, be equivalent in the unit mass dried frozen aquatic products containing propofol 1g, the usage ratio scope of flux oil is at 12-35g.Arbitrary consumption within the scope of weight content all ensures to make stable fatty microemulsion pharmaceutical injection lyophilized formulations.
Embodiment 3: the weight ratio of propofol and synthetic fatty glyceride is 1 ﹕ 12
Prescription:
Get propofol 10.0g tripalmitin 120.0g, add 12.5g DSPC, heated and stirred is about 10-20min mixing.Separately get water for injection 700ml, add mannitol 22.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH6.0-8.0, and filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Get after propofol fat micro emulsion frozen preparation prepared by said method redissolves, adopt supercentrifugation to analyze the content of the free propofol in aqueous phase.
Embodiment 4: the weight ratio of propofol and synthetic fatty glyceride is 1 ﹕ 20
Prescription:
Get propofol 10.0g tripalmitin 200.0g, add 12.5g DSPC, heated and stirred is about 10-20min mixing.Separately get water for injection 600ml, add mannitol 22.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH6.0-8.0, and filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Get after propofol fat micro emulsion frozen preparation prepared by said method redissolves, adopt supercentrifugation to analyze the content of the free propofol in aqueous phase.
Embodiment 5: the weight ratio of propofol and synthetic fatty glyceride is 1 ﹕ 30
Prescription:
Get propofol 10.0g, tripalmitin 300.0g, add 12.5g DSPC, heated and stirred is about 10-20min mixing.Separately get water for injection 600ml, add mannitol 22.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH 6.0-8.0, and filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Get after propofol fat micro emulsion frozen preparation prepared by said method redissolves, adopt supercentrifugation to analyze the content of the free propofol in aqueous phase.
Embodiment 6: the weight ratio of propofol and synthetic fatty glyceride is 1 ﹕ 35
Prescription:
Get propofol 10.0g, tripalmitin 350.0g, add 12.5g DSPC, heated and stirred is about 10-20min mixing.Separately get water for injection 600ml, add mannitol 22.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH 6.0-8.0, and filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Get after propofol fat micro emulsion frozen preparation prepared by said method redissolves, adopt supercentrifugation to analyze the content of the free propofol in aqueous phase.
(2), flux oil amount ranges is investigated
Ensureing that under the prerequisite that the ratio of propofol and flux oil does not change, the amount ranges of the present invention to listed diluent oil is verified.Under basic technology condition of the present invention, the arbitrary consumption of consumption within the scope of 5-35% weight content of listed diluent oil all ensures the novel propofol fat emulsion formulation can making stable painless, low injection stimulation.
Embodiment 7: minimum synthetic fatty acid glyceride content is 5% (weight content).
Prescription:
Get propofol 2.5g, tripalmitin 50.0g, add 12.5g DSPC, heated and stirred is about 10-20min mixing.Separately get water for injection 600ml, add mannitol 22.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH 6.0-8.0, and filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Embodiment 8: synthetic fatty acid glyceride content is 10.0% (weight content).
Prescription:
Get propofol 10g, tripalmitin 100.0g, add 12.5g DSPC, heated and stirred is about 10-20min mixing.Separately get water for injection 600ml, add mannitol 22.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH 6.0-8.0, and filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Embodiment 9: synthetic fatty acid glyceride content is 30.0% (weight content).
Prescription:
Get propofol 16.5g, tripalmitin 300.0g, add 12.5g DSPC, heated and stirred is about 10-20min mixing.Separately get water for injection 600ml, add mannitol 22.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH 6.0-8.0, and filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Embodiment 10: the highest synthetic fatty acid glyceride content is 35.0% (weight content).
Prescription:
Get propofol 16.5g, tripalmitin 350.0g, add 12.5g DSPC, heated and stirred is about 10-20min mixing.Separately get water for injection 600ml, add mannitol 22.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH 6.0-8.0, and filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
(3), the investigation of emulsifying agent synthetic phospholipid consumption
Under the prerequisite that the ratio and flux oil consumption that ensure propofol and flux oil do not change, the amount ranges of the present invention to listed emulsifying agent synthetic phospholipid is verified.Under basic technology condition of the present invention, the arbitrary consumption of consumption within the scope of 0.8-3.2% weight content of listed emulsifying agent all ensures the novel propofol fat emulsion formulation can making stable painless, low injection stimulation.
Embodiment 11: minimum synthetic phospholipid use amount content is 0.8% (weight content)
Prescription:
Get propofol 10.0g, tripalmitin 250.0g, add 8.0g DSPC, heated and stirred is about 10-20min mixing.Separately get water for injection 600ml, add mannitol 22.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH 6.0-8.0, filters, fill, uses brown bottle, prepares lyophilized formulations.
Embodiment 12: synthetic phospholipid use amount content is 1.25% (weight content)
Prescription:
Get propofol 10.0g, tripalmitin 250.0g, add 12.5g DSPC, heated and stirred is about 10-20min mixing.Separately get water for injection 600ml, add mannitol 22.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH 6.0-8.0, and filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Embodiment 13: synthetic phospholipid use amount content is 2.5% (weight content)
Prescription:
Get propofol 10.0g, tripalmitin 250.0g, add 25g DSPC, heated and stirred is about 10-20min mixing.Separately get water for injection 600ml, add mannitol 22.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH 6.0-8.0, and filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Embodiment 14: the highest synthetic phospholipid use amount content is 3.2% (weight content)
Prescription:
Get propofol 10.0g, tripalmitin 250.0g, add 32g DSPC, heated and stirred is about 10-20min mixing.Separately get water for injection 600ml, add mannitol 22.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH 6.0-8.0, and filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
(4) investigation of different skeletal support agent, is used
Embodiment 15: lactose is 2% (weight content) as skeletal support agent use amount
Prescription:
Get propofol 10.0g, glyceryl linolenate 250.0g, add 12.5g DPPC, heated and stirred is about 10-20min mixing.Separately get water for injection 600ml, add lactose 20.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in lactose aqueous solution, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH6.0-8.0, and filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Embodiment 16: lactose is 5% (weight content) as skeletal support agent use amount
Prescription:
Get propofol 10.0g, tristerin 250.0g, add 12.5g linolenic acid phosphatidylcholine, heated and stirred is about 10-20min mixing.Separately get water for injection 600ml, add lactose 50.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in lactose aqueous solution, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH6.0-8.0, and filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Embodiment 17: maltose is 2% (weight content) as skeletal support agent use amount
Prescription:
Get propofol 10.0g, olein 250.0g, add 12.5g linoleic acid phosphatidylcholine, heated and stirred is about 10-20min mixing.Separately get water for injection 600ml, add maltose 20.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in maltose solution, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH6.0-8.0, and filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Embodiment 18: maltose is 5% (weight content) as skeletal support agent use amount
Prescription:
Get propofol 10.0g, glyceryl linoleate 250.0g, add 12.5g DPPC, heated and stirred is about 10-20min mixing.Separately get water for injection 600ml, add maltose 50.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in maltose solution, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH 6.0-8.0, and filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Embodiment 19: mannitol is 2% (weight content) as skeletal support agent use amount
Prescription:
Get propofol 10.0g, tripalmitin 250.0g, add 12.5g DSPC, heated and stirred is about 10-20min mixing.Separately get water for injection 600ml, add mannitol 20.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH 6.0-8.0, and filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Embodiment 20: mannitol is 5% (weight content) as skeletal support agent use amount
Prescription:
Get propofol 10.0g, tripalmitin 250.0g, add 12.5g DSPC, heated and stirred is about 10-20min mixing.Separately get water for injection 600ml, add mannitol 50.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH 6.0-8.0, and filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Method three, use polyethyleneglycol lipid derivates are to the investigation reducing free propofol content in the rear aqueous phase of preparation redissolution
Embodiment 21: the propofol fat micro emulsion frozen preparation adding 1% polyethyleneglycol lipid derivates (with weight note)
Prescription:
Get propofol 10.0g, tripalmitin 200.0g, add 12.5g DSPC and 10g (Polyethylene Glycol-DSPE) DSPE-PEG, heated and stirred is about 10-20min mixing.Separately get water for injection 800ml, add mannitol 22.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, and than the fat emulsion formulation of routine, its particle diameter reduces to some extent, and to particle size range at 50nm-120nm, regulate pH 6.0-8.0, filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Embodiment 22: the propofol fat micro emulsion frozen preparation adding 5% polyethyleneglycol lipid derivates (with weight note)
Prescription:
Get propofol 10.0g, tripalmitin 200.0g, add 12.5g DSPC and 50g (Polyethylene Glycol-DSPE) DSPE-PEG, heated and stirred is about 10-20min mixing.Separately get water for injection 800ml, add mannitol 22.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, and than the fat emulsion formulation of routine, its particle diameter has obvious reduction, and to particle size range at 50nm-120nm, regulate pH 6.0-8.0, filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Embodiment 23: the propofol fat micro emulsion frozen preparation adding 10% polyethyleneglycol lipid derivates (with weight note)
Prescription:
Get propofol 10.0g, tripalmitin 200.0g, add 12.5g DSPC and 100g (Polyethylene Glycol-DSPE) DSPE-PEG, heated and stirred is about 10-20min mixing.Separately get water for injection 800ml, add mannitol 22.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, and than the fat emulsion formulation of routine, its particle diameter has obvious reduction, and to particle size range at 50nm-120nm, regulate pH 6.0-8.0, filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Embodiment 24: the propofol fat micro emulsion frozen preparation adding 15% polyethyleneglycol lipid derivates (with weight note)
Prescription:
Get propofol 10.0g, tripalmitin 200.0g, add 12.5g DSPC and 150g (Polyethylene Glycol-DSPE) DSPE-PEG, heated and stirred is about 10-20min mixing.Separately get water for injection 800ml, add mannitol 22.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, and than the fat emulsion formulation of routine, its particle diameter has obvious reduction, and to particle size range at 50nm-120nm, regulate pH 6.0-8.0, filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Method four, lyophilized formulations use the investigation of the light durability of colourless cillin bottle and brown cillin bottle
Embodiment 25: use colourless bottle and brown bottle on the impact of lyophilized formulations stability
Prescription:
Get propofol 10.0g, tripalmitin 200.0g, add 12.5g DSPC, heated and stirred is about 10-20min mixing.Separately get water for injection 600ml, add mannitol 22g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 100ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH6.0-8.0, and filter, fill, half uses colourless cillin bottle, and half uses brown cillin bottle, prepares lyophilized formulations.
The anaphylaxis adding micro-soybean protein or yolk protein in method five, novel formulation is investigated
Embodiment 26: add micro-soybean protein in novel formulation, anaphylaxis is investigated
Prescription:
Get propofol 10.0g, tripalmitin 250.0g, add 12.5g DSPC, soybean protein 4mg, heated and stirred is about 10-20min mixing.Separately get water for injection 600ml, add mannitol 22.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH 6.0-8.0, and filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Embodiment 27: add micro-yolk protein in novel formulation, anaphylaxis is investigated
Prescription:
Get propofol 10.0g, tripalmitin 250.0g, add 12.5g DSPC, yolk protein 4mg, heated and stirred is about 10-20min mixing.Separately get water for injection 600ml, add mannitol 22.0g.Under nitrogen protection and stirring condition, propofol phosphatide oil solution is added in Osmitrol, regulate total amount to 1000ml.Through high pressure homogenizer homogenizing 7-8 time, homogenization pressure is 100MPa, is less than 180nm to particle diameter, regulates pH 6.0-8.0, and filter, fill, uses brown cillin bottle, prepare lyophilized formulations.
Propofol fat micro emulsion frozen preparation prepared by various method, the comparison of anaphylaxis and reduction injection pain
According to " after redissolving, in water, free propofol content measures ", " granularity and particle size distribution method " and " the irritated inspection technique of propofol fat micro emulsion frozen preparation " inspection in aforementioned.All embodiments are checked, after product redissolves, the free propofol aqueous phase content in solution all≤4 μ g/ml, propofol envelop rate does not reduce and particle diameter is all less than 180nm meets the requirements.In the result that anaphylaxis checks, the sodium chloride injection matched group of 0.9% is attacked in last administration for 10 days, Cavia porcellus is all normal, occur without any anaphylaxis, the Ovum Gallus domesticus album normal saline group of 5% is attacked in last administration for 10 days, first Cavia porcellus occurs grabbing nose, trembles, erects hair, dyspnea, tic, urinary incontinence, finally dead.Death mostly occurred in 2 minutes.Commercially available Propofol fat emulsion injection heavy dose group appearance one example 2 order reaction, grabs nose, trembles, erects hair, cough phenomenon.The administration of self-control propofol fat micro emulsion frozen preparation (embodiment 1-25) last is attacked all without any symptom, without anaphylaxis for 10 days.Embodiment 26,27 is because deliberately adding micro-soybean protein and yolk protein, and last administration is attacked for 10 days, and Cavia porcellus occurs grabbing nose, trembles, erects hair, dyspnea, tic, urinary incontinence, finally dead.Death mostly occurred in 2 minutes.Fatty micro emulsion frozen preparation prepared by visible use prescription of the present invention and compound method can improve the anaphylaxis of former preparation, becomes without anaphylaxis novel formulation.
According to the specific descriptions of aforementioned " important analysis method " neutralization " evaluation test of propofol fat micro emulsion frozen preparation vascular pain ", give propofol fat micro emulsion frozen sample that commercially available and embodiment prepared the voluntarily femoral artery to rat respectively, evaluated the degree of vascular pain by the electromyography at the contiguous position of the tremulous pulse of administration sample.Electromyography record is started before administration propofol fat sample.To Post operation 1 hour, electromyography waveform stabilization, by the intubate embodiment sample prepared of the commercially available propofol fat emulsion of administration and the embodiment of the present invention respectively, record electromyogram respectively after administration, and calculate area under the peak in electromyogram.The value measuring each group of every rat compares with reference to medicine with commercially available, to determine its " electromyogram area ratio ", represents with (%), represents the index of vascular pain.When this index is less than 100%, it shows that vascular pain alleviates compared with commercially available propofol fat emulsion injection.And index (electromyogram area ratio) is less, it is better that vascular pain alleviates effect.
Under the Electromyographic peak of Fig. 6, invention laboratory sample method 1-4 area and commercial samples electromyogram peak under the ratio of area.
1), the sample (embodiment 1) prepared of hyperfiltration process;
2) sample (embodiment 14) prepared by phospholipid consumption, is increased;
3) sample (embodiment 24), using polyethyleneglycol lipid derivates to prepare;
4) sample (embodiment 10) of oil preparation at high proportion, is used.
Check according to " not adding EDTA, sulphite, the propofol fat micro-emulsion injecta of thiosulfate and comparing of lyophilized formulations germ contamination " in aforementioned, observed result is visible:
Without colony growth in the culture dish of inoculation C; In the culture dish of inoculation A or B, extension rate is larger, the bacterial population of the interior growth of culture dish is fewer; In the culture medium inoculating the A of equal extension rate, total number of bacteria is obviously more than the culture dish of inoculation B.Visible sample preparation is become lyophilized formulations after, obviously can reduce bacterial growth.
According to " Propofol fat emulsion injection quality standard " to method 7, use the preparation of the lyophilizing of colourless bottle and use brown bottle, carry out influence factor's exposure experiments to light investigation of 5 days and 10 days.
As seen from the experiment, it is more stable than using colourless cillin bottle properties of samples to use brown cillin bottle.Because propofol contains phenolic hydroxyl group, easily oxidized, adopt brown bottle effectively can prevent the degraded of propofol
[12].
Comprehensive the above results, the propofol phenol fat micro emulsion frozen preparation using the present invention to prepare effectively eliminates possible sensitization source, without anaphylaxis; Decrease the propofol that after redissolving, aqueous phase is free, reduce vascular stimulation, significantly alleviate intravascular injection pain, wherein ultrafiltration, Polyethylene Glycol reduces the effect of pain clearly.It is to be noted that the sample of ultrafiltration can remove pain substantially especially.The method (increasing the consumption of phospholipid or oil) promoting the solubility property of propofol in solvent also can play certain effect, but effect is obvious not as good as directly removing free drug.Sample is made lyophilized formulations and solve the difficult problem not having antioxidant to protect the oxidative degradation of lower propofol and adjuvant; Light degradation when using brown cillin bottle effectively can reduce storage.By said method, we can obtain without sensitization, without the novel propofol fat micro emulsion frozen preparation of injection pain, and make said preparation safety higher.
List of references:
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C,LESSARD M R.Propofol and the risk of transmission ofinfection[J].Can J Anaesth,2003,50:533-537.
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Claims (12)
1., without sensitization, a painless propofol fat micro emulsion frozen preparation, it is characterized in that, composed of the following components:
2. propofol fat micro emulsion frozen preparation according to claim 1, it is characterized in that, described synthetic phospholipid is the mixing of one or more of formula I structure;
R
2:-CH
2-CH
2-N
+H
3(PE)
-CH
2-CH
2-N
+(CH
3)
3(PC)
-C
6H(OH)
5(PI)
-CH
2COO
-CH(N
+H
3) (PS)
-CH
2CHOH-CH
2OH (PG)
Wherein,
R
2represent and be selected from choline (PC), ethanolamine (PE), inositol (PI), glycerol (PG), serine (PS), R and R
1can be identical or different, be selected from saturated, cholesterol or polyunsaturated in, long chain hydrocarbon groups, as alkyl, thiazolinyl, alkynyl, R and R
1length optional 4 ~ 26 carbon of carbochain, the length of preferred carbochain is 6 ~ 22 carbon, and more preferably the length of carbochain is 10 ~ 20 carbon.
3. propofol fat micro emulsion frozen preparation according to claim 1, it is characterized in that, described synthetic fatty glyceride is the mixing of one or more of formula II structure;
Wherein, R, R
1and R
2for can be identical or different, be selected from saturated, cholesterol or polyunsaturated middle long chain hydrocarbon groups, as alkyl, thiazolinyl, alkynyl, R, R
1and R
2length optional 4 ~ 26 carbon of carbochain, the length of preferred carbochain is 6 ~ 22 carbon, and more preferably the length of carbochain is 10 ~ 20 carbon.
4. propofol fat micro emulsion frozen preparation according to claim 1, it is characterized in that, described co-emulsifier is the mixing of one or more in oleic acid, oleate or synthetic mono fatty acid glyceride; Described synthetic mono fatty acid glyceride is the mixing of one or more of formula III structure;
Wherein, R is selected from saturated, cholesterol or polyunsaturated middle long chain hydrocarbon groups, and as alkyl, thiazolinyl, alkynyl, the length of the carbochain of R is 16 ~ 18 carbon.
5. propofol fat micro emulsion frozen preparation according to claim 1, it is characterized in that, described skeletal support agent is one or more in lactose, sucrose, maltose, mannitol, sorbitol, xylitol.
6. propofol fat micro emulsion frozen preparation according to claim 1, it is characterized in that, the weight ratio of propofol and synthetic fatty glyceride is less than 1:15.
7. propofol fat micro emulsion frozen preparation according to claim 1, it is characterized in that, the weight ratio of propofol, synthetic fatty glyceride and synthetic phospholipid is 1:25:(0.8 ~ 3.2).
8. propofol fat micro emulsion frozen preparation according to claim 1, is characterized in that, composed of the following components:
9. the method for preparation propofol fat micro emulsion frozen preparation according to any one of claim 1-7, it is characterized in that, whole process is carried out under nitrogen protection, and comprises
Get heated and stirred, the mixings such as the propofol of recipe quantity, synthetic phospholipid, synthetic fatty glyceride and Polyethylene Glycol phospholipid derivative, separately get appropriate water for injection, add oleic acid and skeletal support agent is dissolved,
Under nitrogen protection condition, oil solution is added slowly in the aqueous phase of cutter high-speed stirred (8000rpm), continue 15min; colostrum solution, through high pressure homogenizer homogenizing 7 ~ 8 times, pressure 100MPa; regulate pH6.0-8.0, size controlling is being less than 180nm
Adopt the independent of multiple method and conbined usage to reach reduce or substantially remove the propofol that in microemulsion aqueous phase, free propofol and emulsion droplet outer surface stick, filter, fill,
Prepare lyophilized formulations: at subzero less than 45 DEG C pre-freeze 2 ~ 5h, be warmed up between subzero 30 DEG C to subzero 15 DEG C and maintain more than 22h, vacuum decompression removing moisture, be warming up to 0 ~ 5 DEG C, maintain 5h and continue vacuum drying decompression removing moisture, be warming up to 25 DEG C, continue vacuum decompression and go out moisture, obtain dry freeze-dried emulsion.
10. preparation method according to claim 9, it is characterized in that: after fatty microemulsion homogenizing completes, adopt the independent of multiple method and conbined usage to reach and reduce or substantially remove the propofol that in the rear aqueous phase of micro emulsion frozen preparation redissolution, free propofol and emulsion droplet outer surface stick.
11. preparation methoies according to claim 9, is characterized in that: the sample particle diameter after high pressure homogenization is less than 180nm.
12. preparation methoies according to claim 9, is characterized in that: select brown cillin bottle fill.
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| CN (1) | CN104523591B (en) |
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| CN105287406A (en) * | 2015-11-17 | 2016-02-03 | 西安力邦肇新生物科技有限公司 | Propofol liposome freeze-drying preparation and preparation method thereof |
| CN106214668A (en) * | 2016-07-21 | 2016-12-14 | 西安力邦制药有限公司 | Propofol flexible nano-liposomes patch and application thereof |
| CN108254450A (en) * | 2016-12-29 | 2018-07-06 | 上海医药集团股份有限公司 | In Propofol/detection method of long chain fat emulsion injection envelop rate |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105287406A (en) * | 2015-11-17 | 2016-02-03 | 西安力邦肇新生物科技有限公司 | Propofol liposome freeze-drying preparation and preparation method thereof |
| CN105287406B (en) * | 2015-11-17 | 2018-12-04 | 西安力邦肇新生物科技有限公司 | A kind of Propofol lipidosome freeze-dried preparation and preparation method thereof |
| CN106214668A (en) * | 2016-07-21 | 2016-12-14 | 西安力邦制药有限公司 | Propofol flexible nano-liposomes patch and application thereof |
| CN108254450A (en) * | 2016-12-29 | 2018-07-06 | 上海医药集团股份有限公司 | In Propofol/detection method of long chain fat emulsion injection envelop rate |
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| CN104523591B (en) | 2019-01-18 |
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